Superstrata microstate geometries furnish some of the most successful laboratories, to date, for probing black hole microstructure in a geometric setting. This paper extends the (1,m,n) family of superstrata, to allow for flat asymptotics. Previous constructions utilized the decoupling regime, where the geometry is asymptotically AdS3xS3. Brief comments are made on the additional complexity introduced by the flat space coupling, how this obscures holomorphicity and breaks the consistent truncation to which the decoupled solutions belong. Holomorphicity and consistent truncation were key simplifications assisting previous studies of the (1,m,n) superstrata, undertaken in the decoupling regime. Further, these results open a window for future projects to determine how previous analysis on the decoupled geometries extend or are modified once flat space asymptotics are imposed. Since our universe is almost flat on cosmological scales, this represents progress towards more phenomenologically relevant microstate geometries. This work can be considered a continuation of that in arxiv.org/abs/1711.10474 where some single mode superstrata were also coupled to flat space.

Heart valve disease is a growing problem worldwide. Though very common in older adults, the mechanisms behind the development of the disease aren’t well understood, and at present the only therapeutic option is valve replacement. Valvular interstitial cells (VICs) may hold the answer. These cells can undergo pathological differentiation into contractile myofibroblasts or osteoblasts, leading to thickening and calcification of the valve tissue. Our study aimed to characterise the effect of fibroblast growth factor 2 (FGF-2) on the differentiation potential of VICs. We isolated VICs from diseased human valves and treated these cells with FGF-2 and TGF-β to elucidate effect of these growth factors on several myofibroblastic outcomes, in particular immunocytochemistry and gene expression. We used TGF-β as a positive control for myofibroblastic differentiation. We found that FGF-2 promotes a ‘quiescent-type’ morphology and inhibits the formation of α-smooth muscle actin positive myofibroblasts. FGF-2 reduced the calcification potential of VICs, with a marked reduction in the number of calcific nodules. FGF-2 interrupted the ‘canonical’ TGF-β signalling pathway, reducing the nuclear translocation of the SMAD2/3 complex. The panel of genes assayed revealed that FGF-2 promoted a quiescent-type pattern of gene expression, with significant downregulations in typical myofibroblast markers α smooth muscle actin, extracellular matrix proteins, and scleraxis. We did not see evidence of osteoblast differentiation: neither matrix-type calcification nor changes in osteoblast associated gene expression were observed. Our findings show that FGF-2 can reverse the myofibroblastic phenotype of VICs isolated from diseased valves and inhibit the calcification potential of these cells.

Research in heart valve biology is a growing field that has yet to elucidate the fundamentals of valve disease. Human valvular interstitial cells (hVICs) are the best option for studying the cellular mechanisms behind valvular pathologies. However, there is a wide range of isolation procedures for these cells published in the literature. To what extent various isolation methods, patient pathologies, and seeding densities influence the behaviour of hVICs remains unclear. Here, we present an optimised method of hVIC isolation from diseased human valves donated at the time of surgery. We show that two rounds of 1000 U/mL collagenase digestion for not >2 h results in a phenotypically stable cell culture with a near complete absence of endothelial cell contamination. We also suggest that cells should be seeded at 10,000 cells/cm2 for experimentation. We found that patient pathology does not affect the success of the isolation procedure, and that instead, successful cultures are predicted by ensuring >500 mg valve tissue as starting material.

Tissue engineered heart valves may one day offer an exciting alternative to traditional valve prostheses. Methods of construction vary, from decellularised animal tissue to synthetic hydrogels, but the goal is the same: the creation of a 'living valve' populated with autologous cells that may persist indefinitely upon implantation. Previous failed attempts in humans have highlighted the difficulty in predicting how a novel heart valve will perform in vivo . A significant hurdle in bringing these prostheses to market is understanding the immune reaction in the short and long term. With respect to innate immunity, the chronic remodelling of a tissue engineered implant by macrophages remains poorly understood. Also unclear are the mechanisms behind unknown antigens and their effect on the adaptive immune system. No silver bullet exists, rather researchers must draw upon a number of in vitro and in vivo models to fully elucidate the effect a host will exert on the graft. This review details the methods by which the immunogenicity of tissue engineered heart valves may be investigated and reveals areas that would benefit from more research. Statement of significance Both academic and private institutions around the world are committed to the creation of a valve prosthesis that will perform safely upon implantation. To date, however, no truly non-immunogenic valves have emerged. This review highlights the importance of preclinical immunogenicity assessment, and summarizes the available techniques used in vitro and in vivo to elucidate the immune response. To the authors knowledge, this is the first review that details the immune testing regimen specific to a TEHV candidate. (c) 2021 The Authors. Published by Elsevier Ltd on behalf of Acta Materialia Inc. This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by-nc-nd/4.0/ )

Abstract Background Recent reports have described the contribution of adult respiratory syncytial virus (RSV) infections to the use of advanced healthcare resources and death. Methods Data regarding patients aged ≥18 years admitted to any of Maryland’s 50 acute-care hospitals were evaluated over 12 consecutive years (2001–2013). We examined RSV and influenza (flu) surveillance data from the US National Respiratory and Enteric Virus Surveillance System and the Centers for Disease Control and Prevention and used this information to define RSV and flu outbreak periods in the Maryland area. Outbreak periods consisted of consecutive individual weeks during which at least 10% of RSV and/or flu diagnostic tests were positive. We examined relationships of RSV and flu outbreaks to occurrence of 4 advanced medical outcomes (hospitalization, intensive care unit admission, intubated mechanical ventilation, and death) due to medically attended acute respiratory illness (MAARI). Results Occurrences of all 4 MAARI-related hospital advanced medical outcomes were consistently greater for all adult ages during RSV, flu, and combined RSV–flu outbreak periods compared to nonoutbreak periods and tended to be greatest in adults aged ≥65 years during combined RSV–flu outbreak periods. Rate ratios for all 4 MAARI-related advanced medical outcomes ranged from 1.04 to 1.38 during the RSV, flu, or combined RSV–flu outbreaks compared to the nonoutbreak periods, with all 95% lower confidence limits >1. Conclusions Both RSV and flu outbreaks were associated with surges in MAARI-related advanced medical outcomes (hospitalization, intensive care unit admission, intubated mechanical ventilation, and death) for adults of all ages.

BACKGROUND:Since 1999, the US Food and Drug Administration approved neuraminidase and endonuclease inhibitors to treat uncomplicated outpatient influenza but not severe hospitalized influenza. After the 2009 pandemic, several influenza hospital-based clinical therapeutic trials were unsuccessful, possibly due to certain study factors. Therefore, in 2014, the US Health and Human Services agencies formed a Working Group (WG) to address related clinical challenges.METHODS:Starting in 2014, the WG obtained retrospective data from failed hospital-based influenza therapeutic trials and nontherapeutic hospital-based influenza studies. These data allowed the WG to identify factors that might improve hospital-based therapeutic trials. These included primary clinical endpoints, increased clinical site enrollment, and appropriate baseline enrollment criteria.RESULTS:During 2018, the WG received retrospective data from a National Institutes of Health hospital-based influenza therapeutic trial that demonstrated time to resolution of respiratory status, which was not a satisfactory primary endpoint. The WG statisticians examined these data and believed that ordinal outcomes might be a more powerful primary endpoint. Johns Hopkins' researchers provided WG data from an emergency-department (ED) triage study to identify patients with confirmed influenza using molecular testing. During the 2013-2014 influenza season, 4 EDs identified 1074 influenza-patients, which suggested that triage testing should increase enrollment by hospital-based clinical trial sites. In 2017, the WG received data from Northwestern Memorial Hospital researchers regarding 703 influenza inpatients over 5 seasons. The WG applied National Early Warning Score (NEWS) at patient baseline to identify appropriate criteria to enroll patients into hospital-based therapeutic trials.CONCLUSIONS:Data received by the WG indicated that hospital-based influenza therapeutic trials could use ordinal outcome analyses, ED triage to identify influenza patients, and NEWS for enrollment criteria.

Clinical Trials Registration:ClinicalTrials.gov [NCT02378753] and Pan African Clinical Trials Registry [PACTR201502001037220].

The West Africa Ebola epidemic stimulated rapid implementation of Ebola vaccine trials in the 3 highly affected countries. In Sierra Leone, we studied the recombinant vesicular stomatitis virus Ebola vaccine (rVSV Delta G-ZEBOV-GP) safety and efficacy. The Sierra Leone Trial to Introduce a Vaccine against Ebola (STRIVE) was a randomized, unblinded Phase 2/3 trial with phased vaccine introduction, no placebo, and concurrent evaluation of vaccine safety and efficacy. Healthcare and frontline response workers in 5 districts were randomized to immediate or deferred (18-24 weeks later) vaccination and followed for 6 months postvaccination. We enrolled 8651 participants from April through August 2015; 7998 were vaccinated. No participants developed Ebola virus disease so an efficacy assessment was not possible. Overall, 132 (1.5%) participants experienced serious adverse events (SAEs); none were vaccine-related. In a detailed safety substudy (N=436), vaccinated participants reported significantly more systemic adverse events (AEs) within 7 days than unvaccinated participants including fever higher than 38 degrees C (20.5% vs 3.9%), headache (71.2% vs 22.1%), fatigue (50.7% vs 10.4%), and joint pain (31.7% vs 6.5%); most AEs were mild to moderate severity and resolved within 5 days. During days 5-28, vaccinated participants more commonly reported joint pain (17.0% vs 4.8%) and rash (7.8% vs 1.7%) (P<.05 for both comparisons). Vaccinated participants also more commonly reported skin vesicles (2.0% vs 0%) and mouth ulcers (2.0% vs 0%) but only during days 8-14 (P<.05 for both comparisons). Among almost 8000 high-risk workers vaccinated during the Sierra Leone Ebola epidemic, rVSV Delta G-ZEBOV-GP was generally well tolerated with no vaccine-related SAEs. Reported joint pain, rash, skin vesicles, and mouth ulcers postvaccination are consistent with conditions associated with transient viral replication described among participants in other trials.

Tuberculosis (TB) is the leading cause of infectious death worldwide. Development of improved TB vaccines that boost or replace BCG is a major global health goal. ID93 + GLA-SE is a fusion protein TB vaccine candidate combined with the Toll-like Receptor 4 agonist adjuvant, GLA-SE. We conducted a phase 1, randomized, double-blind, dose-escalation clinical trial to evaluate two dose levels of the ID93 antigen, administered intramuscularly alone or in combination with two dose levels of the GLA-SE adjuvant, in 60 BCG-naive, QuantiFERON-negative, healthy adults in the US (ClinicalTrials.gov identifier: NCT01599897). When administered as 3 injections, 28 days apart, all dose levels of ID93 alone and ID93 + GLA-SE demonstrated an acceptable safety profile. All regimens elicited vaccine-specific humoral and cellular responses. Compared with ID93 alone, vaccination with ID93 + GLA-SE elicited higher titers of ID93-specific antibodies, a preferential increase in IgG1 and IgG3 subclasses, and a multifaceted Fc-mediated effector function response. The addition of GLA-SE also enhanced the magnitude and polyfunctional cytokine profile of CD4(+) T cells. The data demonstrate an acceptable safety profile and indicate that the GLA-SE adjuvant drives a functional humoral and T-helper 1 type cellular response.

Experiments described in this work examine degradation mechanisms of nickel-based anodes in solid oxide fuel cells (SOFCs) operating with a biogas surrogate and exposed to 110 ppm Cl (delivered either as CH3Cl or HCl).

Time-resolved fluorescence and differential scanning calorimetry were used to determine the partitioning of coumarin 152 (C152) into large unilamellar vesicles composed of binary mixtures of two phosphatidylcholines (12:0/12:0 DLPC and 14:0/14:0 DMPC) and vesicles composed of binary mixtures of a phosphatidylcholine and a phosphatidylethanolamine (14:0/14:0 DMPC and 14:0/14:0 DMPE). Differential scanning calorimetry showed that both DLPC/DMPC and DMPC/DMPE are miscible in lipid vesicles. Time-resolved fluorescence indicated that C152 partitioning into DLPC/DMPC mixtures showed nearly ideal behavior that was described with weighted contributions from C152 partitioning into pure DLPC and pure DMPC vesicles. In contrast, C152 partitioning into DMPC/DMPE mixtures was distinctly nonideal. For DMPC/DMPE lipid vesicles having DMPC mole fractions between 10 and 80%, C152 partitioning into the bilayer was measurably enhanced near the melting temperature, relative to expectations based simply on weighted contributions from C152 partitioning into vesicles comprised of pure lipids. The origin of this behavior remains uncertain. For vesicles comprised of pure DMPE, C152 shows almost no partitioning into the membrane, with ≥80% of the solute remaining in the buffer solution at temperatures between 10 and 50 °C.

Partitioning of coumarin 152 (C152) in phosphatidylcholine vesicles was quantified using time-resolved fluorescence emission. Phospholipid vesicles were comprised of 1,2-dilauroyl-sn-glycero-3-phosphocholine (12:0 DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (14:0 DMPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (16:0 DPPC). C152 fluorescence emission decays were fit to three lifetimes, corresponding to C152 solvated by the aqueous buffer, embedded in polar lipid headgroups, and surrounded by the nonpolar lipid membrane core. Partitioning was measured as a function of sample temperature and vesicle composition. C152 in all three lipid systems showed qualitatively similar partitioning behavior. Partitioning into a gel phase membrane was thermoneutral and slightly entropically favored. Partitioning of C152 near the lipid membrane headgroups was entropically driven and endothermic. Well above the melting temperature, exsolvation of C152 from the membrane back into the aqueous buffer was enthalpically driven but entropically unfavorable. Regardless of solution temperature, relatively little (<15%) C152 partitions into the hydrophobic core of the membrane. The magnitudes of the forces driving C152 partitioning systematically increased with alkyl chain length (DLPC < DMPC < DPPC). Differences in partitioning between the three vesicle systems are attributed to differences in surface area per lipid as membrane phase changes from the gel to liquid-crystalline state.

Background: We report a first-in-human trial evaluating safety and immunogenicity of a recombinant BCG, AERAS-422, over-expressing TB antigens Ag85A, Ag85B, and Rv3407 and expressing mutant perfringolysin.Methods: This was a randomized, double-blind, dose-escalation trial in HIV-negative, healthy adult, BCG naive volunteers, negative for prior exposure to Mth, at one US clinical site. Volunteers were randomized 2:1 at each dose level to receive a single intradermal dose of AERAS-422 (>10(5)-< 10(6) CELL low dose, >= 10(6)- < 10(7) CFU = high dose) or non recombinant Tice BCG (1-8 x 10(5) CFU). Randomization used an independently prepared randomly generated sequence of treatment assignments. The primary and secondary outcomes were safety and immunogenicity, respectively, assessed in all participants through 182 days post-vaccination. ClinicalTrials.gov registration number: NCT01340820.Findings: Between Nov 2010 and Aug 2011, 24 volunteers were enrolled (AERAS-422 high dose, n = 8; AERAS-422 low close, n = 8; Tice BCG, n = 8); all were included in the safely and immunogenicity analyses. All 24 subjects had at least one adverse event, primarily expected local reactions. High close AERAS-422 vaccination induced Ag85A- and Ag85B-specific lymphoproliferative responses and marked anti-mycobacterial activity in a whole blood bactericidal activity culture assay (WBA), but was associated with varicella zoster virus (VZV) reactivalion in two vaccinees. These volunteers displayed high BCG-specific IEN-gamma responses pre- and post-vaccination possibly predisposing them to autocrinelparacrine negative regulation of immune control of latent VZV. A systems biology transcriptomal approach identified positive correlations between post-vaccination T cell expression modules and WBA, and negative correlations between post-vaccination monocyte expression modules and WBA The expression of one key macrophage marker (F4/80) was constitutively elevated in the two volunteers with zoster. Interpretation: The unexpected development of VZV in two of eight healthy adult vaccine recipients resulted in discontinuation of AERAS-422 vaccine development. Immunological and transcriptomal data identified correlations with the development of TB immunity and VZV that require further investigation.Funding: Aeras, FDA, Bill and Melinda Gates Foundation. (C) 2016 The Authors Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Despite widespread use of BCG, tuberculosis (TB) remains a global threat, with 9 million new cases and 1.5 million deaths reported in 2013. Current vaccine candidates in clinical trials are designed to replace BCG or boost the variable efficacy observed with BCG. One of these is AERAS-402, a replication-deficient Ad35 vaccine encoding a fusion protein of the M. tuberculosis antigens 85A, 85B, and TB10.4. This vaccine has been tested in adults, infants, HIV+ adults and patients on or following treatment for TB and shown to be safe and immunogenic. A phase 2 clinical trial of two doses of AERAS-402 was conducted in healthy infants in South Africa, Kenya, and Mozambique. The original study design consisted of a dose-finding phase followed by an efficacy phase at the selected dose. The trial design was modified to include a third dose of AERAS-402. AERAS-402 was well-tolerated at all dose levels and up to three doses. Despite the addition of a third dose, the predefined immunogenicity target was not met and the efficacy portion of the trial was removed. The third dose failed to increase immune responses, which consisted of a dominant polyfunctional (IFN-γ, IL-2, TNF) and bifunctional (IFN-γ, TNF) CD8 response and a lower magnitude polyfunctional and bifunctional (IL-2, TNF) CD4 responses. These responses were considerably lower than those observed in adults. Alternative methods to enhance responses to AERAS-402, such as heterologous prime-boost approaches, are warranted.

Background: New tuberculosis (TB) vaccines are being developed to combat the global epidemic. A phase IIb trial of a candidate vaccine, MVA85A, was conducted in a high burden setting in South Africa to evaluate proof-of-concept efficacy for prevention of TB in infants.Objective: To describe the study design and implementation lessons from an infant TB vaccine efficacy trial.Methods: This was a randomised, controlled, double-blind clinical trial comparing the safety and efficacy of MVA85A to Candin control administered to 4-6-month-old, BCG-vaccinated, HIV-negative infants at a rural site in South Africa. Infants were followed up for 15-39 months for incident TB disease based on pre-specified endpoints.Results: 2797 infants were enrolled over 22 months. Factors adversely affecting recruitment and the solutions that were implemented are discussed. Slow case accrual led to six months extension of trial follow up.Conclusion: The clinical, regulatory and research environment for modern efficacy trials of new TB vaccines are substantially different to that when BCG vaccine was first evaluated in infants. Future infant TB vaccine trials will need to allocate sufficient resources and optimise operational efficiency. A stringent TB case definition is necessary to maximize specificity, and TB case accrual must be monitored closely. (C) 2013 Elsevier Ltd. All rights reserved.

In this paper we describe the architecture of MTASC, a multithreaded associative SIMD processor, and a cycle-accurate instruction set simulator for that architecture. We show, through simulations of a set of five associative benchmarks, that this architecture is capable of significantly improving the performance of associative code over a single-threaded architecture, especially for processors with a large number of PEs. Furthermore, we show that the amount of improvement in performance is highly dependent on the frequency of reduction instructions in the code being executed.

BACKGROUND:Live-attenuated influenza vaccine (LAIV) prevents more cases of influenza in immune-competent children than the trivalent inactivated vaccine (TIV). We compared the antibody responses to LAIV or TIV in HIV-infected children.METHODS:Blood and saliva obtained at enrollment, 4 and 24 weeks postimmunization from 243 HIV-infected children randomly assigned to TIV or LAIV were analyzed.RESULTS:Both vaccines increased the anti-influenza neutralizing antibodies at 4 and 24 weeks postimmunization. At 4 weeks postimmunization, TIV recipients had 2-fold to 3-fold higher neutralizing antibody titers than LAIV recipients, but the proportions of subjects with protective titers (≥ 1:40) were similar between treatment groups (96%-100% for influenza A and 81%-88% for influenza B). Both vaccines increased salivary homotypic IgG antibodies, but not IgA antibodies. Both vaccines also increased serum heterosubtypic antibodies. Among HIV-specific characteristics, the baseline viral load correlated best with the antibody responses to either vaccine. We used LAIV-virus shedding as a surrogate of influenza infection. Influenza-specific humoral and mucosal antibody levels were significantly higher in nonshedders than in shedders.CONCLUSIONS:LAIV and TIV generated homotypic and heterosubtypic humoral and mucosal antibody responses in HIV-infected children. High titers of humoral or mucosal antibodies correlated with protection against viral shedding.